Improving Baculovirus Infectivity by Efficiently Embedding Enhancing Factors into Occlusion Bodies
Improving Baculovirus Infectivity by Efficiently Embedding Enhancing Factors into Occlusion Bodies
复制标题
通过将增强因子有效嵌入闭塞体中提高杆状病毒感染性
DOI：
10.1128/aem.00595-17
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发表时间：
2017-05
影响因子：
4.4
通讯作者：
Sun Xiulian
中科院分区：
文献类型：
--
作者：
Yang Shili;Zhao Lijuan;Ma Ruipeng;Fang Wei;Hu Jia;Lei Chengfeng;Sun Xiulian
摘要杆状病毒对宿主幼虫的相对较低的感染性限制了它们作为杀虫剂的使用。在本研究中，制定了一种新的策略，以有效地将外国蛋白质嵌入加利福尼亚的Autographa
ABSTRACT The relatively low infectivity of baculoviruses to their host larvae limits their use as insecticidal agents on a larger scale. In the present study, a novel strategy was developed to efficiently embed foreign proteins into Autographa californica multiple nucleopolyhedrovirus (AcMNPV) occlusion bodies (OBs) to achieve stable expression of foreign proteins and to improve viral infectivity. A recombinant AcMNPV bacmid was constructed by expressing the 150-amino-acid (aa) N-terminal segment of polyhedrin under the control of the p10 promoter and the remaining C-terminal 95-aa segment under the control of the polyhedrin promoter. The recombinant virus formed OBs in Spodoptera frugiperda 9 cells, in which the occlusion-derived viruses were embedded in a manner similar to that for wild-type AcMNPV. Next, the 95-aa polyhedrin C terminus was fused to enhanced green fluorescent protein, and the recombinant AcMNPV formed fluorescent green OBs and was stably passaged in vitro and in vivo. The AcMNPV recombinants were further modified by fusing truncated Agrotis segetum granulovirus enhancin or truncated Cydia pomonella granulovirus ORF13 (GP37) to the C-terminal 95 aa of polyhedrin, and both recombinants were able to form normal OBs. Bioactivity assays indicated that the median lethal concentrations of these two AcMNPV recombinants were 3- to 5-fold lower than that of the control virus. These results suggest that embedding enhancing factors in baculovirus OBs by use of this novel technique may promote efficient and stable foreign protein expression and significantly improve baculovirus infectivity. IMPORTANCE Baculoviruses have been used as bioinsecticides for over 40 years, but their relatively low infectivity to their host larvae limits their use on a larger scale. It has been reported that it is possible to improve baculovirus infectivity by packaging enhancing factors within baculovirus occlusion bodies (OBs); however, so far, the packaging efficiency has been low. In this article, we describe a novel strategy for efficiently embedding foreign proteins into AcMNPV OBs by expressing N- and C-terminal (dimidiate) polyhedrin fragments (150 and 95 amino acids, respectively) as fusions to foreign proteins under the control of the p10 and polyhedrin promoters, respectively. When this strategy was used to embed an enhancing factor (enhancin or GP37) into the baculovirus OBs, 3- to 5-fold increases in baculoviral infectivity were observed. This novel strategy has the potential to create an efficient protein expression system and a highly efficient virus-based system for insecticide production in the future.
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DOI：
--
发表时间：
2003
期刊：
Chinese journal of virology
影响因子：
--
作者：
Zhong Jiang
通讯作者：
Zhong Jiang
影响因子：
4.4
作者：
H. Shim;J. Choi;Yong Wang;X. Tao;Qin Liu;J. Roh;Jae Su Kim;W. Kim;S. Woo;B. Jin;Y. Je
通讯作者：
H. Shim;J. Choi;Yong Wang;X. Tao;Qin Liu;J. Roh;Jae Su Kim;W. Kim;S. Woo;B. Jin;Y. Je
苹果颗粒病毒GP37对核多角体病毒感染力和苏云金芽孢杆菌致死率的协同作用
影响因子：
2.7
作者：
Liu, Xiangyang;Ma, Xiaohe;Lei, Chengfeng;Xiao, Yuzhou;Zhang, Zhongxin;Sun, Xiulian
通讯作者：
Sun, Xiulian
DOI：
--
发表时间：
1992
期刊：
--
影响因子：
--
作者：
D. O'reilly;L. Miller;V. Luckow
通讯作者：
D. O'reilly;L. Miller;V. Luckow
影响因子：
4.8
作者：
Gramkow AW;Perecmanis S;Sousa RL;Noronha EF;Felix CR;Nagata T;Ribeiro BM
通讯作者：
Ribeiro BM